Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures
Abstract Several laboratories have explored the capacity of the anion exchange chromatography method in evaluating distinct forms of inositol phosphates in a single analytical process. We describe a straightforward HPLC method to analyze simultaneously inositol phosphates and nucleotides present in...
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Wiley
2025-06-01
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| Series: | Physiological Reports |
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| Online Access: | https://doi.org/10.14814/phy2.70419 |
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| author | Natalie Chaves Ferreira Chiara Gatnau‐Civardi Miquel Riera‐Codina |
| author_facet | Natalie Chaves Ferreira Chiara Gatnau‐Civardi Miquel Riera‐Codina |
| author_sort | Natalie Chaves Ferreira |
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| description | Abstract Several laboratories have explored the capacity of the anion exchange chromatography method in evaluating distinct forms of inositol phosphates in a single analytical process. We describe a straightforward HPLC method to analyze simultaneously inositol phosphates and nucleotides present in a neuronal cells culture. The method was applied to neuroblastoma cells grown in standard media. The culture has an optimal metabolic state between 45% and 95% of confluence, but there was a rapid metabolic deterioration when the culture density exceeded 100%, which is important to consider when cell stimulation studies are carried out in culture. This method also allows the quantification since 0.5 to 50 nanomoles of nucleotides present in a single confluent culture from a T‐75 flask containing 8 million cells. In addition, cyclic adenosine monophosphate (cAMP) and adenosine monophosphate (AMP) were eluted without overlapping. Therefore, the method has proven to have sufficient sensitivity to determine quantitative changes in nucleotides and inositol phosphates in a sample with low cell density. Moreover, the simultaneous determination of signaling and metabolic molecules allows obtaining a rapid and suitable control of the metabolic status in studies on cell stimulation that should be applicable to other types of cultured cells. |
| format | Article |
| id | doaj-art-42b13931bb48451b87013bb44894b97a |
| institution | Kabale University |
| issn | 2051-817X |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | Physiological Reports |
| spelling | doaj-art-42b13931bb48451b87013bb44894b97a2025-08-20T03:28:01ZengWileyPhysiological Reports2051-817X2025-06-011312n/an/a10.14814/phy2.70419Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell culturesNatalie Chaves Ferreira0Chiara Gatnau‐Civardi1Miquel Riera‐Codina2Department of Cellular Biology, Physiology and Immunology, Faculty of Biology University of Barcelona Barcelona 08028 SpainDepartment of Cellular Biology, Physiology and Immunology, Faculty of Biology University of Barcelona Barcelona 08028 SpainDepartment of Cellular Biology, Physiology and Immunology, Faculty of Biology University of Barcelona Barcelona 08028 SpainAbstract Several laboratories have explored the capacity of the anion exchange chromatography method in evaluating distinct forms of inositol phosphates in a single analytical process. We describe a straightforward HPLC method to analyze simultaneously inositol phosphates and nucleotides present in a neuronal cells culture. The method was applied to neuroblastoma cells grown in standard media. The culture has an optimal metabolic state between 45% and 95% of confluence, but there was a rapid metabolic deterioration when the culture density exceeded 100%, which is important to consider when cell stimulation studies are carried out in culture. This method also allows the quantification since 0.5 to 50 nanomoles of nucleotides present in a single confluent culture from a T‐75 flask containing 8 million cells. In addition, cyclic adenosine monophosphate (cAMP) and adenosine monophosphate (AMP) were eluted without overlapping. Therefore, the method has proven to have sufficient sensitivity to determine quantitative changes in nucleotides and inositol phosphates in a sample with low cell density. Moreover, the simultaneous determination of signaling and metabolic molecules allows obtaining a rapid and suitable control of the metabolic status in studies on cell stimulation that should be applicable to other types of cultured cells.https://doi.org/10.14814/phy2.70419cell metabolismHPLCinositol phosphatesnucleotidessignaling molecules |
| spellingShingle | Natalie Chaves Ferreira Chiara Gatnau‐Civardi Miquel Riera‐Codina Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures Physiological Reports cell metabolism HPLC inositol phosphates nucleotides signaling molecules |
| title | Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| title_full | Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| title_fullStr | Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| title_full_unstemmed | Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| title_short | Simultaneous HPLC determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| title_sort | simultaneous hplc determination of soluble signaling molecules and metabolic status in neuroblastoma cell cultures |
| topic | cell metabolism HPLC inositol phosphates nucleotides signaling molecules |
| url | https://doi.org/10.14814/phy2.70419 |
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